UPLC-MS/MS method for determination of sofosbuvir in human plasma.
| Autor: | Gandhi BM; Department of Pharmaceutical Analysis, Sri Vasavi Institute of Pharmaceutical Sciences, Tadepalligudem, AP, India., Rao AL; Department of Pharmaceutical Analysis, V. V. Institute of Pharmaceutical Sciences, Gudlavalleru, AP, India. Electronic address: dralrao@gmail.com., Rao JV; Department of Pharmaceutical Analysis, St. Paul's College of Pharmacy, Turkayamjal, Hyderabad, TS, India. |
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| Jazyk: | angličtina |
| Zdroj: | Annales pharmaceutiques francaises [Ann Pharm Fr] 2017 Jul; Vol. 75 (4), pp. 257-266. Date of Electronic Publication: 2017 May 08. |
| DOI: | 10.1016/j.pharma.2017.04.003 |
| Abstrakt: | Introduction: A sensitive and rapid method for quantitation of Sofosbuvir in human plasma has been established using ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). Materials and Methods: Sofosbuvir d3 was used as an internal standard. Sofosbuvir and internal standard in plasma sample were extracted using ethyl acetate (liquid liquid extraction). A centrifuged upper layer was then evaporated and reconstituted with the mobile phase of 0.5% formic acid: methanol (30:70, v/v). The reconstituted samples were injected into a Gemini C18 (50×4.6mm, 5μm) column. Results: Using MS/MS in the multiple reaction monitoring mode, Sofosbuvir and Sofosbuvir d3 were detected without severe interferences from human plasma matrix. Sofosbuvir produced a protonated precursor ion ([M+H] + ) at m/z 428.35 and a corresponding product ion at m/z 279.26. The internal standard produced a protonated precursor ion ([M+H] + ) at m/z 431.38 and a corresponding product ion at m/z 282.37. The calibration curves for the analyte was linear (R 2 ≥0.9956, n=4) over the concentration range of 4.063-8000.010ng/mL. Stability studies revealed that Sofosbuvir was stable in plasma during bench top (7h at room temperature), in injector (20h), at the end of five successive freeze and thaw cycles and long term at -70°C±15°C for 15 days. Conclusion: The developed method was validated as per the guidelines of USFDA and the obtained results were found to be within the limits and could be successfully employed for the determination of Sofosbuvir in human plasma for regular and pharmacokinetic studies. (Copyright © 2017 Académie Nationale de Pharmacie. Published by Elsevier Masson SAS. All rights reserved.) |
| Databáze: | MEDLINE |
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